SEQUENCING TUMOR SUPPRESSOR LOCI

描述(申请人的描述) 我们的目标是利用基因组测序和分析来识别 我们推测的候选肿瘤抑制基因存在于 我们已经发现在人类癌症中存在纯合缺失。我们将：a) 确定该病毒完整和准确的核苷酸序列 在人类癌症中发现重复缺失的染色体区域， 特别强调乳腺癌；b)使用序列分析和 数据库搜索以从这些区域中寻找候选基因；c) 确定有效肿瘤的完整内含子/外显子结构 从这些区域转录的抑制基因。 目前正在对两个区域进行测序：区域 携带PTEN的10号染色体，PTEN是一种肿瘤抑制基因 在晚期乳腺癌、脑癌和前列腺癌中经常发生突变；以及 20号染色体上在胃肠道中经常缺失的区域 肿瘤。多个基因座正在鉴定过程中，包括 8号染色体上的两个位置，一个在21号染色体上，一个在4号染色体上，它们被删除了 在各种癌症中。我们预计将对以下内容进行排序和分析 每年5个基因座，每个约300-400kb。 这项工作将补充冷泉正在进行的其他工作 使用RDA(代表性差异)的港湾癌症中心 分析)以确定缺失的基因座，使用外显子捕获和 杂交选择从这些基因座中识别候选基因，并使用 突变分析以验证候选基因是否参与 临床癌症。

DESCRIPTION (Applicant's Description) Our objective is to utilize genomic sequencing and analysis to identify candidate tumor suppressors that we presume reside within regions that we have found are homozygously deleted in human cancers. We shall: a) determine the complete and accurate nucleotide sequence of the chromosomal regions found to be repeatedly deleted in human cancers, with a special emphasis on breast cancer; b) use sequence analysis and data base searching to find candidate genes from these regions; c) determine the complete intron/exon structures for the validated tumor suppressor genes transcribed from these regions. Sequencing of two regions is currently in progress: the region on chromosome 10 that harbors PTEN, a tumor suppressor gene that is frequently mutated in advanced breast, brain and prostate cancers; and a region on chromosome 20 that is frequently deleted in gastrointestinal tumors. Multiple loci are in the process of characterization, including two sites on chromosome eight, one on 21, and one on 4 that are deleted in a variety of cancers. We expect to be sequencing and analyzing about 5 loci per year, each about 300-400 kb in length. This work will complement other work in progress at the Cold Spring Harbor Cancer Center that utilizes RDA (representational difference analysis) to identify deleted loci, uses methods of exon trapping and hybrid selection to identify candidate genes from these loci, and uses mutational analysis to validate the involvement of candidate genes in clinical cancer.